PRODUCT
SpCas9 (H840A)-NLS
Cat#
CASPT-017
Source Organism
Streptococcus pyogenes
Expression Host Organism
E. coli
Tag Type
His
Sequence
full length
Introduction
Recombinant full-length SpCas9 (H840A)-NLS from S. pyogenes expressed in E. coli using a C-terminal His tag. Contains a nuclear localization signal for optimal activity. Enzyme sold with 1 mL of Cas9 reaction buffer, 10x (CS09-09).
Size
50 ug/100 ug/250 ug/1000 ug/Bulk Customize
Alternate Names
Cas9-H840A Nickase Protein
UniProt Number
Q99ZW2
Overview
Cas9 is a multidomain nuclease from the bacterium Streptococcus pyogenes characteristic of type II CRISPR-Cas systems (1). Guided by a single-guide RNA (sgRNA), Cas9 targets specific genomic loci and generates a blunt-ended double-strand break (2). SpCas9 (H840A)-NLS is a catalytic mutant of Cas9 containing a nuclear localization signal for optimal activity (3). Unlike the wild-type Cas9, SpCas9 (H840A)-NLS generates a single-strand DNA break on the non-targeting strand. SpCas9 (H840A)-NLS can be used for various genome editing applications.
Accession Number
AAK33936.1
Formulation
Protein stored in 50 mM sodium phosphate, pH 7.5, 300 mM NaCl, 200 mM imidazole, 1 mM DTT, 10% glycerol
References
1. Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O. Crystal structure of Cas9 in complex with guide RNA and target DNA. 2014. Cell. 156(5):935-49.
2. Sapranauskas R, Gasiunas G, Fremaux C, Barrangou R, Horvath P, Siksnys V. 2011. The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli. Nucleic Acids Res. 39(21):9275-82.
3. Jiang F, Doudna JA. 2017. CRISPR-Cas9 Structures and Mechanisms. Annu Rev Biophys.46:505-529.
2. Sapranauskas R, Gasiunas G, Fremaux C, Barrangou R, Horvath P, Siksnys V. 2011. The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli. Nucleic Acids Res. 39(21):9275-82.
3. Jiang F, Doudna JA. 2017. CRISPR-Cas9 Structures and Mechanisms. Annu Rev Biophys.46:505-529.
Storage and Stability
Store at –70°C. To avoid repeated handling and multiple freeze/thaw cycles, aliquot product into smaller quantities.
Concentration
0.2 μg/μl
6.26 pmol/μg
6.26 pmol/μg
Purity
>70%
Molecular weight
Calculated: 161.0 kDa
Observed: ~160 kDa
Observed: ~160 kDa
Recommendation
Always include RNase inhibitor at recommended concentrations in Cas enzyme reactions.
